Allogeneic Immunotherapy for cancer and nonmalignant hematological disorders

癌症和非恶性血液病的同种异体免疫疗法

• 批准号: 8557917
• 负责人: RICHARD CHILDS
• 金额: $ 260.95万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8557917
• 关键词: Accounting; Activated Natural Killer Cell; Adoptive Immunotherapy; Adoptive Transfer; Adult; Aftercare; Allogenic; Allografting; Animal Model; Animals; Antigen Targeting; Antigens; Antithymoglobulin; Aplastic Anemia; Area; Autologous; Back; Blood specimen; Bortezomib; CD34 gene; CSF3 gene; Cancer Patient; Cancer Vaccines; Carcinoma; Cell Line; Cells; Cellular Immunity; Cessation of life; Chimerism; Clear Cell; Clinical; Clinical Research; Clinical Trials; Correlative Study; Cyclophosphamide; Cytoplasmic Granules; Cytotoxic T-Lymphocytes; DNA; Data; Depsipeptides; Development; Diagnosis; Disadvantaged; Disease; Disseminated Malignant Neoplasm; Donor person; Dose; Effector Cell; Engraftment; Enrollment; Equus caballus; Future; Goals; Graft Rejection; Graft vs Tumor Effect; HERVs; Hematologic Neoplasms; Hematological Disease; Hematopoietic Stem Cell Transplantation; Histone Deacetylase Inhibitor; Homologous Transplantation; Hour; Human; Hypothyroidism; Hypoxia; Immunity; Immunosuppressive Agents; In Vitro; Infusion procedures; Laboratories; Laboratory Research; Ligands; Macaca mulatta; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Mediating; Metastatic Renal Cell Cancer; Methods; Minor; Modeling; Monoclonal Antibodies; Multiple Myeloma; Multivariate Analysis; Mus; Natural Killer Cells; Neoplasm Metastasis; Neutropenia; Non-Malignant; Normal tissue morphology; Pancytopenia; Patients; Pharmaceutical Preparations; Phenotype; Population; Proteins; Proviruses; Recovery; Refractory; Regimen; Renal Cell Carcinoma; Renal carcinoma; Reporting; Research; Research Personnel; Safety; Secondary to; Small Interfering RNA; Solid Neoplasm; Source; Specificity; Speed; Stem cell transplant; Stem cells; Surface; Survival Rate; Syndrome; System; T-Lymphocyte; TNFSF10 gene; Techniques; Technology; Testing; Therapeutic; Thyroiditis; Toxic effect; Translating; Translational Research; Transplantation; Tumor Antigens; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; VHL Gene Inactivation; Whole-Body Irradiation; Work; base; cancer cell; cancer immunotherapy; cell killing; chronic graft versus host disease; cohort; conditioning; cytokine; cytotoxic; cytotoxicity; demethylation; expression cloning; fludarabine; follow-up; gene induction; graft vs host disease; improved; in vivo; inhibitor/antagonist; insight; killings; knock-down; leukemia; melanoma; multicatalytic endopeptidase complex; neoplastic cell; neutrophil; novel; overexpression; peripheral blood; phase 1 study; prevent; programs; rat Piga protein; receptor; research study; selective expression; transcription factor; tumor

Approximately 10 years ago, we initiated the first clinical trail investigating allogeneic stem cell transplantation in patients with treatment refractory metastatic renal cell carcinoma (RCC). Definitive evidence for an allogeneic graft-versus-RCC effect has been demonstrated with complete regression of large metastasis observed in some patients. We have subsequently conducted studies investigating the immunological mechanisms accounting for tumor regression in patients demonstrating an anti-RCC effect in attempts to identify both the effector cell populations mediating these regressions as well as their target antigens. We demonstrated that cytotoxic T-cell clones with specificity for minor antigens are capable of killing RCC cells in patients having a GVT effect post transplant. Using c-DNA expression cloning, we have recently identified a novel tumor antigen derived from a human endogenous retrovirus over-expressed in RCC cells called HERV-E CT-RCC. This antigen is not expressed on normal tissues and therefore could potentially serve as a target for a future kidney cancer vaccine. Work to identify other antigens derived from this HERV-E that are expressed on common HLA molecules as well as efforts to develop a monoclonal antibody that recognizes HERV-E derived proteins that are selectively expressed in RCC are ongoing. We have also sought to characterize the mechanisms accounting for selective expression of the CT-RCC HERV-E in RCC. Recently, we found that inactivation of the VHL tumor suppressor gene, over-expression of the hypoxia-inducible transcription factor (HIF) HIF-2, and demethylation of the HERV-E 5LTR results in the selective expression of HERV-E in clear cell carcinoma. These studies have provided valuable insights needed for ongoing translational research aimed at boosting human immunity against antigenic components of this provirus. In order to explore methods to enhance GVT effects against RCC, we have developed a murine model of allogeneic HCT. This model demonstrates that adoptively transferred NK cells from MHC matched donors reduce GVHD and mediate a GVT effect against murine RCC prolonging survival. These findings are being translated into a clinical trial that will explore the potential of ex-vivo expanded donor NK cell infusions to reduce GVHD and enhance GVT in patients with RCC and other cancers undergoing allogeneic HCT. We are also exploring methods to sensitize solid tumors to NK cell attack by altering the phenotype of tumor cells through targeted gene induction. Recently we showed that bortezomib and depsipeptide sensitize tumors to NK cell cytotoxity by enhancing NK-cell mediated TRAIL killing. This sensitization appears to overcome NK inhibition that is mediated through KIR-KIR ligand interactions. We have also developed a method to expand by > 4 logs NK cells from healthy donors for adoptive infusion in future NK-cell based adoptive immunotherapy trials. We now have developed a method to expand human NK cells by 100 to 1000 fold in bags using GMP conditions. A phase I study entitled Safety and the anti- tumor effects of escalating doses of adoptively infused ex vivo expanded autologous natural killer cells against metastatic cancers or hematological malignancies sensitized to NK TRAIL cytotoxicity with Bortezomib was initiated in 2008 and is currently accruing patients. With the exception of thyroiditis, infusions of up to 2.5x108 expanded NK cells/kg have been well tolerated and provided early evidence for mediating anti-tumor effects in patients with cancer. We have also initiated NK cell studies in rhesus macaques to evaluate methods to optimize the antitumor function and in vivo persistence of adoptively infused autologous NK cells.We have also recently shown that siRNA inactivation of NKG2A enhances NK cell tumor cytotoxicity in vitro and in vivo in tumor-bearing mice, a finding that may have therapeutic implications for humans withcancer. Finally, we have recently partnered with Dr. Ola Landgren from the NCI to explore KIR blockade with the monoclonal antibody IPH 2101 (Innate Pharma) as a method to enhance autologous NK cell killing of smoldering myeloma. Patients with a confirmed diagnosis of smoldering myeloma will receive IPH2101 (1mg/kg) every other month for 6 cycles. Patients will be restaged monthly pre- and post-treatment. In vitro correlative studies from blood samples collected from patients to measure the effects of KIR blockade on NK cell cytotoxic granule release, NK cell cytotoxicity and NK cell intracellular cytokine secretion against myeloma cell lines will be performed in my laboratory. Using this technique, the percentage change in NK cell cytotoxic granule release and NK cell intracellular cytokine secretion following treatment with IPH2101 compared to baseline will be determined and correlated with clinical evidence for disease activity. This study is actively enrolling patients. My group has continued to investigate methods to improve allogeneic HCT for heavily pretreated patients with bone marrow failure syndromes (BMFS) including paroxysmal nocturnal hemoglobinuria (PNH) and ATG-refractory severe aplastic anemia (SAA). In a cohort of 56 heavily transfused and HLA alloimmunized patients with BMFS undergoing a novel fludarabine-based HCT, graft rejection did not occur and 87% of the patients survived disease free with a median follow-up of 4.5 years. Despite this excellent survival, approximately 70% of patients developed chronic GVHD. In a multivariate analysis, we found that rapid donor T-cell engraftment (d 95% donor T-cell chimerism by day 30) was associated with the development of chronic GVHD (adjusted HR 3.03, p=0.04). To test whether delaying the speed of donor T-cell engraftment will prevent chronic GVHD, we have initiated a new clinical trial for BMFS patients which uses a modified donor allograft containing high doses of G-CSF mobilized CD34+ selected cells combined with a one log reduced dose of non-G-CSF mobilized T-cells. Transplantation using unrelated cord blood (UCB) has been shown to be a reasonable alternative transplant approach for patients with a variety of hematological malignancies and non-malignant hematological disorders who lack an HLA matched related or unrelated stem cell donor. Recent data show leukemia-free survival in adult patients with AML and ALL is comparable to that of recipients receiving fully matched unrelated donor transplants. However, for patients with SAA undergoing UCB transplantation, results have been disappointing. Investigators from the EBMT and Eurocord have reported engraftment rates of only 51% and survival rates of only 38% in SAA recipients of single or dual CB transplants. In order to harness the advantage of UCB availability, to overcome the disadvantage of delayed neutrophil recovery of UCB transplantation, and to provide a back up stem cell source should the CB unit fail to engraft, we have recently initiated a clinical trial that evaluates the co-administration of unrelated umbilical cord blood and a relatively low number of highly purified haploidentical peripheral blood CD34+ cells from a related donor as a method to promote rapid neutrophil recovery. Subjects receive a novel non-myeloablative immunosuppressive conditioning regimen of cyclophosphamide, fludarabine, horse ATG and one dose of total body irradiation (200cGy) followed by an infusion of the allografts. The haploidentical stem cell product is T-cell depleted and enriched for CD34+ cells using the Miltenyi CliniMacs system. To reduce TRM secondary to prolonged neutropenia associated with conventional UCB transplantation, haploidentical CD34+ stem cells are co-infused with a single UCB unit (serologically matched at d 4/6 HLA loci). The primary endpoint will be donor engraftment by day 42. This study is now accruing patients.

大约10年前，我们启动了第一个研究同种异体干细胞移植治疗难治性转移性肾细胞癌（RCC）患者的临床试验。同种异体移植物抗rcc效应的明确证据已被证实，在一些患者中观察到的大转移完全消退。我们随后进行了研究，研究了患者肿瘤消退的免疫学机制，证明了抗rcc的作用，试图确定介导这些消退的效应细胞群及其靶抗原。我们证明了对次要抗原具有特异性的细胞毒性t细胞克隆能够杀死移植后具有GVT效应的患者的RCC细胞。利用c-DNA表达克隆技术，我们最近发现了一种新的肿瘤抗原，它来源于一种在RCC细胞中过表达的人内源性逆转录病毒，称为HERV-E CT-RCC。这种抗原在正常组织中不表达，因此有可能作为未来肾癌疫苗的靶标。目前正在进行的工作是鉴定在普通HLA分子上表达的HERV-E衍生的其他抗原，以及开发识别在RCC中选择性表达的HERV-E衍生蛋白的单克隆抗体。我们还试图描述CT-RCC HERV-E在RCC中选择性表达的机制。最近，我们发现VHL肿瘤抑制基因失活、缺氧诱导转录因子（HIF） HIF-2的过度表达以及HERV-E 5LTR的去甲基化导致了HERV-E在透明细胞癌中的选择性表达。这些研究为正在进行的旨在增强人类对该原病毒抗原成分的免疫力的转化研究提供了有价值的见解。为了探索增强GVT抗RCC作用的方法，我们建立了小鼠同种异体HCT模型。该模型表明，来自MHC匹配供体的过继转移NK细胞可降低GVHD，并介导GVT对小鼠RCC的作用，延长生存期。这些发现正在转化为一项临床试验，该试验将探索体外扩增供体NK细胞输注在RCC和其他接受同种异体HCT的癌症患者中降低GVHD和增强GVT的潜力。